高压二氧化碳杀灭大肠杆菌过程中蛋白质变化研究.pdfVIP

现代食品科技 Modern Food Science and Technology 2014, Vol.30, No.5 高压二氧化碳杀灭大肠杆菌过程中的蛋白质变化研究 杨扬，李欣，饶伟丽，陈丽，张德权 （中国农业科学研究院农产品加工研究所，农业部农产品加工重点实验室，北京100193） 摘要：本研究采用 37 ℃、10~50 MPa 的高压二氧化碳（HPCD ）处理大肠杆菌及其全蛋白溶液30 min，检测菌落总数、表观形 态、蛋白质溶解性、蛋白质组和蛋白质二级结构的变化，研究 HPCD 胁迫下大肠杆菌的蛋白质变化规律。结果表明：HPCD 处理压 强升至30 MPa，菌落总数对数值有显著降低；经不同压强处理，菌体形态没有发生显著变化；与对照组（0.1 MPa ）相比，可溶性蛋 白经20 MPa 的HPCD 处理后含量显著降低，从 1.77 mg/mL 降为0.57 mg/mL，并随压强继续上升逐渐降低；30 MPa 的HPCD 处理 菌液，经双向电泳分离到46 个差异最大的蛋白点，质谱鉴定具代表性的 16 个点，分别涉及细胞骨架、新陈代谢、信号转换、运输与 翻译等功能；蛋白溶液圆二色谱结果显示，对照组中 α-螺旋占 68.04%，β-折叠占 0.33%，随HPCD 处理压强增加，α-螺旋含量显著 降低（P0.05 ）。经20 MPa 的HPCD 处理，β-折叠含量显著增加（P0.05 ），但随压强继续增加含量变化不显著。 关键词：高压二氧化碳；大肠杆菌；溶解性；蛋白质组；二级结构 文章篇号：1673-9078(2014)5-118-124 Analysis of Intracellular Proteins of E. coli Inactivated by High Pressure Carbon Dioxide YANG Yang, LI Xin, RAO Wei-li, CHEN Li, ZHANG De-quan (Institute of Agro-Products Processing Science and Technology, Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-Products Processing, Ministry of Agriculture, Beijing 100193, China) Abstract: The variation of proteins in E. coli after 10~50 MPa HPCD treatment at 37 ℃ for 30 min was investigated in this study. The total bacterial counts, cell structure, protein solubility, proteome and secondary structure of protein were analyzed. The results showed that the logarithm of the total bacterial counts of E. coli after HPCD treatment was significantly decreased when the pressure increased to 30 MPa. The cell structure did not change with different HPCD treatments (10~50 MPa). The soluble protein content significantly decreased from 1.77 mg/mL to 0.57 mg/mL when the extraction of E. coli proteins was treated by 20 MPa HPCD in comparison with control (0.1 MPa treatment). The content of prot